盐酸多柔比星脂质体的制备及体外质量评价

目的：制备盐酸多柔比星脂质体并进行体外质量评价，以期实现自制脂质体质量与市售盐酸多柔比星脂质体注射液Doxil一致，并为其生物体内等效及产业化提供研究基础。方法：采用乙醇注入法结合硫酸铵梯度法制备盐酸多柔比星脂质体，以包封率为评价指标通过单因素筛选和正交试验优化制备工艺，以市售Doxil为参比制剂，对比最优工艺制得的脂质体与市售Doxil的微观结构、粒径大小及分布、Zeta电位、相变温度、包封率等理化指标，考察最优工艺制得的脂质体与市售Doxil在加速条件的体外释放及储存条件下稳定性情况。结果：自制盐酸多柔比星脂质体最优制备工艺为高速剪切速率13 000 r·min^-1，挤出3次，载药孵育温度65℃，载药孵育时间60 min，最优工艺制得的脂质体与市售Doxil外观均为椭圆形，平均粒径分别为（90.81±0.58） nm （n=3）和（89.05±0.66） nm （n=3），分布较为均匀，Zeta电位分别为（-41.9±1.8） mV （n=3）和（-44.9±4.2） mV （n=3），相变温度范围相同，包封率分别为98.0%和97.9%，体外释放较为一致，稳定性好。结论：采用最优工艺制备的盐酸多柔比星脂质体制备工艺可行，与市售Doxil相比，理化指标及体外释放无明显差异，自制脂质体体外质量良好，符合实验预期。

Preparation and in vitro quality evaluation of doxorubicin hydrochloride liposome

OBJECTIVE To prepare doxorubicin hydrochloride liposome and evaluate its quality in vitro,in order to achieve the same quality as self-made liposome and Doxil,a commercially available doxorubicin liposome injection,and to provide a research basis for its in vivo equivalence and industrialization.METHODS Doxorubicin hydrochloride liposomes were prepared by ethanol injection combined with ammonium sulfate gradient method.The encapsulation efficiency was evaluated by single factor screening and orthogonal test.The microstructure,particle size and distribution,zeta potential,phase transition temperature,entrapment efficiency and other physical and chemical indicators of liposomes prepared by the optimal process were compared with those of commercial Doxil.The in vitro release and stability of liposomes prepared by the optimal process and commercial Doxil under accelerated conditions were investigated.RESULTS The optimal preparation process of self-made doxorubicin hydrochloride liposome was as follows:high-speed shear rate of 13 000 r·min^-1,3 times of extrusion,drug-loading incubation temperature 65℃,drug-loading incubation time 60 min.The appearance of both liposomes prepared by the optimal process and commercially available Doxil were oval.The average particle sizes were(90.81±0.58)nm(n=3)and(89.05±0.66)nm(n=3),respectively.The distribution was relatively uniform,and the zeta potentials were(-41.9±1.8)mV(n=3)and(-44.9±4.2)mV(n=3).The phase transition temperature range of both preparations was the same,and the entrapment efficiencies were 98.0% and 97.9%,respectively.The in vitro release was more consistent,and the stability was good.CONCLUSION The preparation process of doxorubicin hydrochloride liposome prepared by the optimal process is feasible.Compared with the commercially available Doxil,there is no significant difference between physical and chemical indicators and in vitro release.The quality of self-made liposome is good in vitro and meets the experimental expectation.